Three new shuttle vectors for heterologous expression in Zymomonas mobilis

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• 作者：Qinghua Cao ; Tao Li ; Huanhuan Shao ; Xuemei Tan ; Yizheng Zhang ; ^{yizzhang@scu.edu.cn" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Expression vector ; Gene expression ; Glucoamylase ; Zymomonas mobilis
• 刊名：Electronic Journal of Biotechnology
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：19
• 期：Complete
• 页码：33-40
• 全文大小：765 K

文摘

Zymomonas mobilis, as a novel platform for bio-ethanol production, has been attracted more attention and it is very important to construct vectors for the efficient expression of foreign genes in this bacterium.

Results

Three shuttle vectors (pSUZM1, pSUZM2 and pSUZM3) were first constructed with the origins of replication from the chromosome and two native plasmids (pZZM401 and pZZM402) of Z. mobilis ZM4, respectively. The three shuttle vectors were stable in Z. mobilis ZM4 and have 3, 32 and 27 copies, respectively. The promoter Ppdc (a), from the pyruvate decarboxylase gene, was cloned into the shuttle vectors, generating the expression vectors pSUZM1(2, 3)a. The codon-optimized glucoamylase gene from Aspergillus awamori combined with the signal peptide sequence from the alkaline phosphatase gene of Z. mobilis was cloned into pSUZM1(2, 3)a, resulting in the plasmids pSUZM1a-GA, pSUZM2a-GA and pSUZM3a-GA, respectively. After transforming these plasmids into Z. mobilis ZM4, the host was endowed with glucoamylase activity for starch hydrolysis. Both pSUZM2a-GA and pSUZM3a-GA were more efficient at producing glucoamylase than pSUZM1a-GA.

Conclusions

These results indicated that these expression vectors are useful tools for gene expression in Z. mobilis and this could provide a solid foundation for further studies of heterologous gene expression in Z. mobilis.